Proof of Y-Loss As Clonal Abnormality in MDS by Comparative Analysis of CD34+ and CD3+ Peripheral Blood Cells,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3800-3800 ◽  
Author(s):  
Christina Ganster ◽  
Friederike Braulke ◽  
Katayoon Shirneshan ◽  
Dietrich Kämpfe ◽  
Uwe Platzbecker ◽  
...  
Keyword(s):  
T Cells ◽  
Y Chromosome ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Fish Analysis ◽  
Peripheral Blood Cells ◽  
Clone Size ◽  
Healthy Men ◽  
Age Related ◽  
Cd34 Cells

Abstract Abstract 3800 Introduction: In an ongoing diagnostic study we are currently following chromosomal anomalies in immunomagnetically enriched CD34+ peripheral blood cells in patients with suspected or proven myelodysplastic syndromes (MDS) at short intervals using fluorescence in situ hybridization (FISH) analysis every two to three months over three years. A loss of the Y chromosome was detected in 4% of these patients, as a single anomaly in 2%. Since it is controversially discussed whether loss of the Y chromosome is an age-related or a clonal event in patients with MDS, we aimed to examine whether a Y-loss is clonal or an age-related event in our patients. Methods: For patients with known Y-loss, we used peripheral blood not only to immunomagnetically enrich clonal CD34+ cells, but also CD3+ T-cells not belonging to the MDS clone. Subsequently, we performed FISH analysis to compare the clone sizes of cells with Y-loss in CD34+ and CD3+ cells. As our laboratory threshold for the FISH probe in CD34+ peripheral blood cells is 5%, we included 18 patients with clone sizes exceeding this threshold in CD34+ cells. The median age of the patients was 76 years (range 62–89). To establish a laboratory threshold for the FISH probe in CD3+ peripheral blood cells, we analyzed T-cells of 25 healthy men with a median age of 27 years (range 19–35). Furthermore, we just initialized an investigation of the laboratory threshold for the FISH probe in CD3+ peripheral blood cells of elder men by measuring the frequency of loss of the Y chromosome in T-cells of this control cohort not suffering from hematopoietic diseases. Until now we could recruit 15 men with a median age of 75 years (range 66–84) for this purpose, further will follow soon. Results: In patients with suspected or proven MDS, the number of cells with -Y was significantly increased in CD34+ cells compared to CD3+ cells (p<0.0001). The median clone size was 64% (range 12–97) in CD34+ cells and 5% (range 1–14) in CD3+ T-cells. The clone size in CD34+ cells was at least four times higher than in CD3+ cells in all patients. We could not detect further chromosomal abnormalities in 16 patients. Chromosomal banding analysis revealed that cells with -Y and cells with -Y and +8 occurred in parallel in two patients. In men below the age of 35 Y-loss could not be detected. The median clone size of 0.5% (range 0–2) resulted in a laboratory threshold of 2%. Interim analysis of men over the age of 65 resulted in a median clone size of 2.5% (range 1–14) and a laboratory threshold of 13%. So far the FISH-signal corresponding to the Y chromosome was significantly more frequent missing in T-cells of elder than in T-cells of younger men (p=0.005). Conclusion: Regarding the absence of Y-loss in CD3+ peripheral blood cells of young healthy men compared to up to 14% -Y in CD3+ peripheral blood cells of elder men, the low proportion of -Y in CD3+ cells of our patients suggests an age related Y-loss in normal T-cells. As the number of CD34+ peripheral blood cells with -Y exceeds the number of CD3+ peripheral blood cells with -Y in all patients, we assume that Y-loss is clonal to some extent in all of them. We established a reliable method to test if loss of the Y chromosome is disease- or age-related in individual MDS patients. It can be used to determine a clonal disease in patients with suspected MDS and Y-loss as sole abnormality. Disclosures: No relevant conflicts of interest to declare.

Loss of the Y Chromosome in MDS - Age-Related Phenomenon or Clonal Abnormality?

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4008-4008
Author(s):  
Christina Ganster ◽  
Friederike Braulke ◽  
Katayoon Shirneshan ◽  
Francesc Solé ◽  
Mar Mallo ◽  
...  
Keyword(s):  
Experimental Study ◽  
T Cells ◽  
Y Chromosome ◽  
Karyotype Evolution ◽  
Normal Karyotype ◽  
Related Phenomenon ◽  
Clone Size ◽  
Age Related ◽  
Cd34 Cells ◽  
First Diagnosis

Abstract Abstract 4008 Introduction: Loss of the Y chromosome has been reported to be associated with hematopoietic diseases (Wiktor et al., 2000), but it was also described as an age-related phenomenon in males (UKCCG, 1992). Determination of clonality and prediction of prognosis and treatment outcome might benefit from a differentiation between age- and MDS-associated Y loss. The aim of this study was to evaluate if Y loss was an age- and/or MDS-associated phenomenon by retrospectively analyzing our multicenter, international DACH-, ICWG- and IMRAW-database and by testing the established hypotheses in an experimental study. Patients and Methods: In our multicenter MDS-database of 2901 patients, 101 primary, untreated MDS patients (3.5%) with loss of the Y were identified. We analyzed them according to age, clone size, and the presence or absence of additional chromosomal aberrations and assessed the prognostic relevance of the aberrations using univariate and multivariate models. Additionally, by immunomagnetic cell sorting, we enriched clonal CD34+ cells and CD3+ T-cells not belonging to the MDS clone from peripheral blood of three patients and compared the percentage of cells with -Y using FISH. Results: Isolated loss of Y was observed in 65.3% (n=66) of the 101 patients identified in the multicenter MDS-database, 14.9% (n=15) of the patients displayed one additional aberration and in 19.8% (n=20) -Y occurred as part of complex abnormalities. Overall survival of patients with -Y as a sole change was significantly better compared to patients with a normal karyotype (60.8 vs. 47.4 months; hazard ratio = 0.50, p<0.01). Loss of the Y chromosome as isolated aberration was significantly less frequent in younger (<60 years) than in older patients (1.9% vs. 4.0%. p<0.01). Patients showing -Y as single aberration were older at time of first diagnosis as patients with any other abnormalities or a normal karyotype (71.5 vs. 66.7 years, p<0.01). There were no differences in clone size between patients with -Y and patients with 5q-, -7/7q-, 20q- or complex karyotypes. Studying sequential karyotypes of some patients, we observed -Y occurring during karyotype evolution. We also identified a patient with -Y at first diagnosis in a mosaic karyotype with normal cells that later developed additional aberrations in the cells with -Y during the course of the disease. These data strongly suggest that -Y is MDS-associated in these patients. To demonstrate that -Y is occurring in the clonal CD34+ cellular compartment as a somatically acquired event we studied the loss of the Y chromosome in CD34+ and CD3+ cells separately in 3 patients. The percentage of cells with -Y was significantly increased in CD34+ cells (69%, 74%, 47%) compared to CD3+ cells (8%, 7%, 2%; p=0.01). Until now, it is not clear whether the low proportion of loss of the Y chromosome in CD3+ cells that exceed in two cases only slightly our laboratory threshold of 3.8% is due to an age related loss of the Y chromosome in T-cells or to contamination of the CD3+ cells with clonal cells. Conclusion: The frequency of loss of Y chromosome in MDS karyotypes is associated with age. However, the evidently better prognosis of MDS patients with loss of the Y chromosome as the sole aberration compared to MDS patients with normal karyotype, the occurrence of Y loss during karyotype evolution, the acquisition of additional abnormalities in cells with -Y as primary aberration, and the observation of -Y in clonal CD34+ cells but not in CD3+ cells suggest a clonal nature of this karyotype anomaly at least in a subset of patients with MDS. In other patients this finding is clearly age-related. The preliminary data of our experimental study will be tested by analyzing a larger cohort of patients. Disclosures: No relevant conflicts of interest to declare.

Receptor Mediated Binding of the Fibrinolytic Components, Plasminogen and Urokinase, to Peripheral Blood Cells

1987 ◽  
Vol 58 (03) ◽  
pp. 936-942 ◽  
Author(s):  
Lindsey A Miles ◽  
Edward F Plow
Keyword(s):  
T Cells ◽  
B Cell ◽  
Peripheral Blood ◽  
Binding Sites ◽  
Blood Cells ◽  
High Capacity ◽  
Red Cells ◽  
Peripheral Blood Cells ◽  
Cellular Functions ◽  
Fibrinolytic Proteins

SummaryGlu-plasminogen binds to platelets; the monocytoid line, U937, and the human fetal fibroblast line, GM1380 bind both plasminogen and its activator, urokinase. This study assesses the interaction of these fibrinolytic proteins with circulating human blood cells. Plasminogen bound minimally to red cells but bound saturably and reversibly to monocytes, granulocytes and lymphocytes with apparent Kd values of 0.9-1.4 μM. The interactions were of high capacity with 1.6 to 49 × 105 sites/cell and involved the lysine binding sites of plasminogen. Both T cells and non-rosetting lymphocytes and two B cell lines saturably bound plasminogen. Urokinase bound saturably to gianulocytes, monocytes, non-rosetting lymphocytes and a B cell line, but minimally to T cells, platelets and red cells. Therefore, plasminogen binding sites of high capacity, of similar affinities, and with common recognition specificities are expressed by many peripheral blood cells. Urokinase receptors are also widely distributed, but less so than plasminogen binding sites. The binding ol plasminogen and/ or urokinase to these cells may lead to generation of cell- associated proteolytic activity which contributes to a variety of cellular functions.

scholarly journals Age-related but not longevity-related genes are found by weighted gene co-expression network analysis in the peripheral blood cells of humans

2018 ◽  
Vol 93 (6) ◽  
pp. 221-228 ◽  
Author(s):  
Chunhong Li ◽  
Dan Mo ◽  
Meiqin Li ◽  
Yanyan Zheng ◽  
Qiao Li ◽  
...  
Keyword(s):  
Network Analysis ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Peripheral Blood Cells ◽  
Age Related

High Transgene Expression Rates After Extended Follow up Among Rhesus Macaque Recipients of Autologous Hematopoietic Stem Cells Transduced with a Modified HIV1-Based Lentiviral Vector

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3118-3118
Author(s):  
Naoya Uchida ◽  
Phillip W. Hargrove ◽  
Coen J. Lap ◽  
Oswald Phang ◽  
Aylin C. Bonifacino ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Transgene Expression ◽  
Vector System ◽  
Large Animal ◽  
Peripheral Blood Cells ◽  
Hematopoietic Stem ◽  
Integration Pattern ◽  
Cd34 Cells ◽  
Integration Sites

Abstract Abstract 3118 Hematopoietic stem cell (HSC)-targeted gene therapy is potentially curative for the hemoglobin disorders; however, highly efficient, lineage specific globin expression remains elusive, and large animal models thus remain important for further development toward clinical application. We previously constructed a chimeric HIV1 vector (χHIV vector) system to circumvent a species specific restriction to HIV1-based vectors wherein the HIV1 vector genome is packaged in the context of the simian immunodeficiency virus (SIV) capsid for efficient transduction of rhesus CD34+ cells in vitro (J Virol. 2009) and in vivo (ASH 2009). In this study, we sought to evaluate transduction efficiency and vector integration pattern among long-term repopulating cells in the rhesus HSC transplantation model. We followed up transgene expression rates among peripheral blood cells of three animals for 1.5–2 years. For two animals (RQ7307 and RQ7280), half of the CD34+ cells were transduced with a standard SIV vector and the other half with the χHIV vector using the same protocol. Transduced cells were transplanted into lethally irradiated rhesus macaques, as previously described (J Virol. 2009). The transgene expression rates in peripheral blood cells plateaued 3–4 months after transplantation and similar transgene expression rates continued in all cell lineages for at least 1.5 years (Figure). The χHIV vector demonstrated that 2–3 fold higher transgene expression rates were seen in granulocytes (RQ7307: 8.6±0.2% vs. 3.1±0.1%, RQ7280: 27.9±0.7% vs. 18.4±0.2%) and RBCs (RQ7307: 3.3±0.1% vs. 0.9±0.0%, RQ7280: 10.0±0.1% vs. 4.0±0.1%), and equivalent transgene expression rates in lymphocytes (RQ7307: 7.8±0.2% vs. 4.5±0.1%, RQ7280: 22.4±0.5% vs. 17.6±0.3%) and platelets (RQ7307: 3.1±0.1% vs. 2.7±0.1%, RQ7280: 12.3±0.2% vs. 16.8±0.2%), compared to the SIV vector. The average vector copy numbers in transduced cells were 4.6–5.7 for the χHIV vector and 1.5–2.0 for the SIV vector in both transplanted animals, evaluated by Southern blot analysis. We then performed transplantation of rhesus CD34+ cells which were transduced with the χHIV vector alone to evaluate transgene expression and vector integration pattern. Transgene expression rates among peripheral blood cells in this animal (RQ7387) plateaued 1–3 months after transplantation, with stable high transgene expression rates of 51.7±1.2% in granulocytes, 54.7±0.1% in lymphocytes, 22.1±0.2% in RBCs, and 19.1±0.1% platelets for 2 years after transplantation. Multi-lineage marking was observed by flow cytometric analysis. We then evaluated integration sites for the χHIV vector in the recipient of χHIV vector alone transduced cells by linear amplification mediated-PCR, using peripheral blood cells of RQ7387 in 0.5–1.5 years after transplantation. We found a total of 344 integration sites for the χHIV vector, and our data demonstrated that the χHIV vector integrated into gene regions, especially introns, when compared to the integration pattern of computer-generated random controls (p<0.001). On the other hand, our data revealed fewer integrations of the χHIV vector into ≤30kb upstream of genes (p<0.001) and into the upstream regions of transcription start sites. Most of the integration sites had low gene density (0–10 genes within 1 Mb upstream or downstream of integration sites, p<0.01), compared to that of random controls. No specific trend was noted for the number of integration sites around CpG islands and the number of CISs around integration sites. These data suggest that the χHIV vector has integration patterns comparable to HIV1 and SIV vectors. In summary, our χHIV vector shows efficient transduction for rhesus long-term repopulating cells, achieving sufficient levels for therapeutic effects in gene therapy trials for globin disorders. This χHIV vector system should allow preclinical testing of HIV1-based therapeutic vectors in large animal models. Disclosures: No relevant conflicts of interest to declare.

In vitro synthesis of antibodies to acetylcholine receptor by peripheral blood cells: Role of suppressor T cells in normal subjects

Neurology ◽  
1984 ◽  
Vol 34 (6) ◽  
pp. 802-802 ◽  
Author(s):  
R. P. Lisak ◽  
C. Laramore ◽  
A. I. Levinson ◽  
B. Zweiman ◽  
A. R. Moskovitz ◽  
...  
Keyword(s):  
T Cells ◽  
Acetylcholine Receptor ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Normal Subjects ◽  
Peripheral Blood Cells ◽  
Suppressor T Cells ◽  
In Vitro Synthesis

Anti-tumor cytotoxicity of γδ T cells expanded from peripheral blood cells of patients with myeloma and lymphoma

2007 ◽  
Vol 25 (2) ◽  
pp. 137-147 ◽  
Author(s):  
Anri Saitoh ◽  
Miwako Narita ◽  
Norihiro Watanabe ◽  
Nozomi Tochiki ◽  
Noriyuki Satoh ◽  
...  
Keyword(s):  
T Cells ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Γδ T Cells ◽  
Peripheral Blood Cells ◽  
Tumor Cytotoxicity

pattern of cytokine secretion by peripheral blood cells of patients with multiple sclerosis in Brazil

2000 ◽  
Vol 6 (5) ◽  
pp. 293-299 ◽  
Author(s):  
Patrícia Mara da Costa ◽  
Clarissa Lin Yasuda ◽  
Silvia M Scagliusi ◽  
Blanca Maria Diaz-Bardales ◽  
Ernane Maciel ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
T Cells ◽  
Inflammatory Cytokines ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Basic Protein ◽  
Chronic Phase ◽  
Peripheral Blood Cells ◽  
Activated T Cells ◽  
Organ Specific

Autoimmune T cells play a key role as regulators and effectors of organ-specific autoimmune disease. In multiple sclerosis (MS), activated T cells specific for myelin components produce a plethora of inflammatory cytokines and mediators that contribute to myelin damage. The production of proinflammatory and regulatory cytokines by peripheral blood cells from patients with active and stable MS and healthy controls were examined. The results show that TNFa production was somewhat elevated in active MS with no significant increase in the level IFNg, whereas in the chronic phase the anti-inflammatory cytokines IL-10 and TGFb increased, accompanied by a reduction in IFNg when stimulated by myelin basic protein.

Increased loss of the Y chromosome in peripheral blood cells in male patients with autoimmune thyroiditis

2012 ◽  
Vol 38 (2-3) ◽  
pp. J193-J196 ◽  
Author(s):  
Luca Persani ◽  
Marco Bonomi ◽  
Ana Lleo ◽  
Simone Pasini ◽  
Fabiola Civardi ◽  
...  
Keyword(s):  
Y Chromosome ◽  
Autoimmune Thyroiditis ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Peripheral Blood Cells ◽  
Male Patients

scholarly journals FUNCTIONAL ACTIVITY OF CD34-POSITIVE CELLS IN CHRONIC MYELOID LEUKEMIA PATIENTS WITH DIFFERENT RESPONSE TO IMATINIB THERAPY

2015 ◽  
Vol 37 (1) ◽  
pp. 70-72 ◽  
Author(s):  
I O Sviezhentseva ◽  
T P Perekhrestenko ◽  
D I Bilko ◽  
A I Gordienko ◽  
M V Diachenko ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Myeloid Leukemia ◽  
Imatinib Therapy ◽  
Peripheral Blood Cells ◽  
Optimal Response ◽  
Cd34 Cells ◽  
Different Response

Introduction: It is believed that the reason of the leukemic clone cell resistance to treatment with tyrosine kinase inhibitors during chronic myeloid leukemia (CML) is mutations in the genome of an early bone marrow progenitor cells that are CD34-positive. Such cells, regardless of treatment, acquire ability to proliferation and differentiation. This leads to the re-expansion of the CD34+ cells. Aim: to determine the CD34 antigen expression in bone marrow and peripheral blood cells in CML patients with different response to imatinib therapy using the results of hematopoietic cells culturing and the data of flow cytometry. Methods: Bone marrow aspirate from 39 patients who were treated with imatinib was studied with cytogenetic, flow cytometry and culture methods in vitro. Results: In patients with an optimal response to imatinib therapy the number of colonies was 1.8 times lower than the number of those in the group of patients with a suboptimal response to therapy. In turn, in patients with failure of imatinib therapy the number of colonies was the highest and was 2.1 times higher than the patients with optimal response. The results of cytometric studies have shown that the number of CD34+ cells in bone marrow was significantly higher compared to the number of CD34+ cells in peripheral blood cells and increased with the acquisition of leukemic cells the resistance to imatinib. There was a direct correlation between the number of colonies and clusters in semisolid agar in vitro and the number of CD34+ cells in the bone marrow of patients. Conclusions: The correlation between the number of CD34+ cells and the number of cell aggregates in semisolid agar in vitro indicates the prognostic value of the method for determining CD34+ cells in the patient bone marrow. The parallel increase of their number in the peripheral blood will allow developing express methods for the detection of individual patient response to imatinib therapy.

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